As a distinct pathological factor and metastatic route of malignant tumors,perineural invasion(PNI)exhibits a close association with the prognosis of several cancer types.In recent years,the clinical significance and underlying mechanisms of PNI in gynecological malignant tumors have increasingly drawn research attention.Currently,it is recognized that PNI is clinicopathologically and prognostically linked to cervical cancer and vulvar cancer.The exploration of its mechanisms encompasses multiple gynecological tumors and numerous molecular pathways,including the CCL2/CCR2 axis,NGF/TrkA pathway,GluR2,among others.Nevertheless,the existing research models are fragmented,lacking systematicity and in-depth analysis,thus necessitating integration.This paper presents a comprehensive review of the occurrence of PNI in gynecologic malignancies,its correlations with clinicopathology and prognosis,its implications for surgical approaches and postoperative adjuvant therapies,as well as the potential molecular mechanisms.By synthesizing the advancements in both clinical and basic research,this review aims to offer a solid theoretical foundation for further exploration of the mechanisms underlying PNI in gynecologic malignancies and for the optimization of clinical therapeutic strategies.

Objective:To evaluate the effectiveness and safety of 3D simultaneous dual fluorescence imaging using indocyanine green(ICG)and methylene blue(MB)in surgeries for malignant uterine tumors.Methods:37 patients who underwent surgery for cervical or endometrial cancer at Guangxi Medical University Cancer Hospital from April 16,2024 to July 19,2024 were included.All patients received simultaneous fluorescence imaging using ICG and MB during surgery.ICG was injected into the cervix for sentinel lymph node(SLN)imaging,and MB was intravenously administered for ureteral imaging.3D fluorescence laparoscopy was used for simultaneous dual fluo-rescence imaging,and the accuracy of localization,signal quality,and clinical effectiveness were assessed using the Likert scale.Adverse events were monitored,and postoperative follow-up was conducted.Results:The SLN visualization rate was 100.0％,and the ureteral visualization rate was 97.3％.The average start time for SLN ima-ging was 4.08±0.64 min after ICG injection,while the average start time for ureteral imaging was 44.86±5.42 min after intravenous MB administration.The main locations of SLN included:internal iliac(13.5％),external iliac(32.4％),and obturator(67.6％).The localization accuracy and signal quality Likert scores of ICG were 4.59±0.45 points and 4.57±0.50 points,respectively;MB were 4.37±0.88 points and 4.28±0.91 points,respectively.There was no ureteral injury during the operation,no urinary fistula after the operation,and no serious adverse e-vents related to the contrast agents were reported.Conclusions:The 3D simultaneous dual fluorescence imaging technique is highly feasible and safe in surgeries for cervical and endometrial cancers.It can effectively locate SLNs and visualize the ureters,simplify the surgical process and reduce difficulty.This technique provides a new auxiliary method for surgeries involving malignant uterine tumors,with promising clinical application prospects.

Objective:To evaluate the effectiveness and safety of 3D simultaneous dual fluorescence imaging using indocyanine green(ICG)and methylene blue(MB)in surgeries for malignant uterine tumors.Methods:37 patients who underwent surgery for cervical or endometrial cancer at Guangxi Medical University Cancer Hospital from April 16,2024 to July 19,2024 were included.All patients received simultaneous fluorescence imaging using ICG and MB during surgery.ICG was injected into the cervix for sentinel lymph node(SLN)imaging,and MB was intravenously administered for ureteral imaging.3D fluorescence laparoscopy was used for simultaneous dual fluo-rescence imaging,and the accuracy of localization,signal quality,and clinical effectiveness were assessed using the Likert scale.Adverse events were monitored,and postoperative follow-up was conducted.Results:The SLN visualization rate was 100.0％,and the ureteral visualization rate was 97.3％.The average start time for SLN ima-ging was 4.08±0.64 min after ICG injection,while the average start time for ureteral imaging was 44.86±5.42 min after intravenous MB administration.The main locations of SLN included:internal iliac(13.5％),external iliac(32.4％),and obturator(67.6％).The localization accuracy and signal quality Likert scores of ICG were 4.59±0.45 points and 4.57±0.50 points,respectively;MB were 4.37±0.88 points and 4.28±0.91 points,respectively.There was no ureteral injury during the operation,no urinary fistula after the operation,and no serious adverse e-vents related to the contrast agents were reported.Conclusions:The 3D simultaneous dual fluorescence imaging technique is highly feasible and safe in surgeries for cervical and endometrial cancers.It can effectively locate SLNs and visualize the ureters,simplify the surgical process and reduce difficulty.This technique provides a new auxiliary method for surgeries involving malignant uterine tumors,with promising clinical application prospects.

Convergence insufficiency intermittent exotropia (CI-IXT) is a subtype of intermittent exotropia.Surgery remains the most commonly used treatment.Improved unilateral lateral rectus recession-medial rectus resection (I-RR), slanted or nonslanted bilateral medial rectus resection (BMRs) and bilateral slanted lateral rectus recession (S-BLRc) are the commonly used modalities for surgical treatment.These treatment modalities have been able to correct distance and near exotropia and simultaneously reduce the near-distance difference.The clinical application of slanted or nonslanted BMRs is limited due to exodrift over time.I-RR and S-BLRc are widely used in clinics for patients with CI-IXT because of better long-term outcomes.A good knowledge of the different indications of each procedure and its appropriate application can lead to good clinical results.This article reviews recent advances in the surgical management of CI-IXT.

Convergence insufficiency intermittent exotropia (CI-IXT) is a subtype of intermittent exotropia.Surgery remains the most commonly used treatment.Improved unilateral lateral rectus recession-medial rectus resection (I-RR), slanted or nonslanted bilateral medial rectus resection (BMRs) and bilateral slanted lateral rectus recession (S-BLRc) are the commonly used modalities for surgical treatment.These treatment modalities have been able to correct distance and near exotropia and simultaneously reduce the near-distance difference.The clinical application of slanted or nonslanted BMRs is limited due to exodrift over time.I-RR and S-BLRc are widely used in clinics for patients with CI-IXT because of better long-term outcomes.A good knowledge of the different indications of each procedure and its appropriate application can lead to good clinical results.This article reviews recent advances in the surgical management of CI-IXT.

As a distinct pathological factor and metastatic route of malignant tumors,perineural invasion(PNI)exhibits a close association with the prognosis of several cancer types.In recent years,the clinical significance and underlying mechanisms of PNI in gynecological malignant tumors have increasingly drawn research attention.Currently,it is recognized that PNI is clinicopathologically and prognostically linked to cervical cancer and vulvar cancer.The exploration of its mechanisms encompasses multiple gynecological tumors and numerous molecular pathways,including the CCL2/CCR2 axis,NGF/TrkA pathway,GluR2,among others.Nevertheless,the existing research models are fragmented,lacking systematicity and in-depth analysis,thus necessitating integration.This paper presents a comprehensive review of the occurrence of PNI in gynecologic malignancies,its correlations with clinicopathology and prognosis,its implications for surgical approaches and postoperative adjuvant therapies,as well as the potential molecular mechanisms.By synthesizing the advancements in both clinical and basic research,this review aims to offer a solid theoretical foundation for further exploration of the mechanisms underlying PNI in gynecologic malignancies and for the optimization of clinical therapeutic strategies.

Human alphaherpesvirus 2 (HSV-2) is the main pathogen resulting human genital herpes, which poses a major threat to the socio-economic development, while there is no effective vaccine. In this study, we developed a novel lipopolyplex (LPP)-delivered mRNA vaccine expressing the HSV-2 envelope glycoprotein gD and evaluated its immunogenicity in mice. The mRNA vaccine was prepared from the genetically modified gD mRNA synthesized in vitro combined with the LPP delivery platform and it was named gD-ORI mRNA. The expression of gD antigen in the mRNA vaccine was validated in vitro by Western blotting and indirect immunofluorescence assay, then the immune responses induced by this mRNA vaccine in mice were evaluated. The immunization with gD mRNA alone induced strong humoral and cellular immune responses in mice. Robust and long-lasting gD-specific IgG antibodies were detected in the mouse serum after booster immunization with gD-ORI mRNA. The immunized mice exhibited a Th1/Th2 balanced IgG response and robust neutralizing antibodies against HSV-2, and a clear dose-response relationship was observed. The gD-specific IgG antibodies were maintained in mice for a long time, up to 18 weeks post-booster immunization. At the same time, multifunctional gD-specific CD4⁺ and CD8⁺ T cells in vaccinated mice were detected by intracellular cytokine staining (ICS). This novel gD-expressing mRNA vaccine delivered by LPP induces strong and long-lasting immune responses in mice post booster immunization and has a promising prospect for development and application. This study provides scientific evidence and reference for the development of a new mRNA vaccine for HSV-2.
Animals ; Herpesvirus 2, Human/genetics* ; Viral Envelope Proteins/genetics* ; Mice ; Herpes Genitalis/immunology* ; RNA, Messenger/immunology* ; Female ; Mice, Inbred BALB C ; Antibodies, Viral/blood* ; mRNA Vaccines/immunology* ; Antibodies, Neutralizing/blood* ; Humans

Objective:To obtain a prevalent respiratory syncytial virus (RSV) clinical isolate in Beijing and analyze the genotype and biological characteristics of the strain.Methods:A nasopharyngeal secretion specimen was collected from a child with RSV infection in Beijing in 2023 and used for viral isolation. Viral nucleic acid was amplified using qRT-PCR. The isolated virus was identified by transmission electron microscopy, indirect immunofluorescence assay, and plaque formation assay. A phylogenetic analysis was conducted based on the whole-genome sequencing results. Virus titers were determined, and replication characteristics were analyzed. The efficacy of the isolated strain for in vitro screening of antiviral drugs was validated. Results:A clinical RSV isolate, named hRSV/C-Tan/BJ 202301, was successfully isolated, which could form syncytia in Hep-2 cells. Spherical, filamentous, and irregular virus particles were observed by electron microscopy. Immunofluorescence detection showed green fluorescence in Hep-2 cells, and plaque assay showed round plaques, which were similar to the Long strain in morphology. Genomic sequence analysis showed that it belonged to ON1 genotype. It exhibited similar cell growth kinetics characteristics with the Long strain and could be used for antiviral drug screening in vitro. Conclusions:In this study, one RSV strain is successfully isolated and identified. The biological characteristics and the phylogenetic relationship of this strain reflect the characteristics of the circulating strains in Beijing, which provides experimental material for RSV vaccine development and antiviral drug screening in China.

Objective:To obtain a prevalent respiratory syncytial virus (RSV) clinical isolate in Beijing and analyze the genotype and biological characteristics of the strain.Methods:A nasopharyngeal secretion specimen was collected from a child with RSV infection in Beijing in 2023 and used for viral isolation. Viral nucleic acid was amplified using qRT-PCR. The isolated virus was identified by transmission electron microscopy, indirect immunofluorescence assay, and plaque formation assay. A phylogenetic analysis was conducted based on the whole-genome sequencing results. Virus titers were determined, and replication characteristics were analyzed. The efficacy of the isolated strain for in vitro screening of antiviral drugs was validated. Results:A clinical RSV isolate, named hRSV/C-Tan/BJ 202301, was successfully isolated, which could form syncytia in Hep-2 cells. Spherical, filamentous, and irregular virus particles were observed by electron microscopy. Immunofluorescence detection showed green fluorescence in Hep-2 cells, and plaque assay showed round plaques, which were similar to the Long strain in morphology. Genomic sequence analysis showed that it belonged to ON1 genotype. It exhibited similar cell growth kinetics characteristics with the Long strain and could be used for antiviral drug screening in vitro. Conclusions:In this study, one RSV strain is successfully isolated and identified. The biological characteristics and the phylogenetic relationship of this strain reflect the characteristics of the circulating strains in Beijing, which provides experimental material for RSV vaccine development and antiviral drug screening in China.

Objective:To study the expression of methyltransferase-like protein 14 (METTL14) in epithelial ovarian cancer and its clinical significance, and to explore the effect of METTL14 expression on the proliferation, invasion and migration of ovarian cancer cells.Methods:Immunohistochemistry (IHC) was used to detect METTL14 expression in tumor tissue samples, and analyze the relationships among METTL14 expression, clinicopathological factors, and prognosis in ovarian cancer. Lentiviral vectors and small interfering RNA (siRNA) were used to up-regulate and down-regulate the METTL14 expression in ovarian cancer cell lines A2780 and SKOV3, respectively. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was used to detect the N6-methyladenosine (m6A) content in ovarian cancer cells. Cell counting kit-8 (CCK-8), wound healing assay, and transwell assay were used to examine the function of METTL14 expression in the cells.Results:(1) The IHC score of METTL14 protein was 6.2±3.7 in 20 samples of ovarian cancer tissues and 3.3±2.5 in 15 samples of normal ovarian tissues, and the difference was statistically significant ( t=-2.64, P=0.012). Among the patients who suffered from ovarian cancer, there were 69 cases with high expression of METTL14 protein (IHC score≥6), accounting for 57.0% (69/121), and the cases with low expression of METTL14 protein (IHC score<6) accounting for 43.0% (52/121). Compared with the patients with low expression of METTL14, the patients with high expression of METTL14 had later stages, higher rates of lymph node metastasis, abdominal metastasis, and more ascite amount. The differences were statistically significant (all P<0.05). The overall survival rate was significantly lower in patients with high METTL14 expression than the low expression ( P=0.009). (2) LC-MS/MS data showed that the relative expression of m6A in A2780 and SKOV3 cells in the lentivirus (LV)-METTL14 group were 0.213±0.024 and 0.181±0.018, which were significantly higher than those in the LV-normal control (NC) group (0.109±0.022 and 0.128±0.020; all P<0.05). While the relative expression of m6A in A2780 and SKOV3 cells in the si-METTL14 group were 0.063±0.012 and 0.069±0.015, which were significantly lower than the expression in si-NC group of 0.108±0.014 and 0.121±0.014 (all P<0.05). CCK-8 assay showed that the absorbance values were significantly lower in the si-METTL14 group compared with the si-NC group at 36, 48, 60 hours (all P<0.05); while were significantly increased in the LV-METTL14 group compared with the LV-NC group at 48, 60 hours (all P<0.01). Scratch wound assays showed that the migration rate of the si-METTL14 group was lower than those of the si-NC group, while the LV-METTL14 group were higher than the LV-NC group by 24 hours, the differences were statistically significant (all P<0.01). Cell migration and invasion were detected by transwell migration and invasion assays. After cultivated for 24 hours, the invasion cell number and the migration cell number in the si-METTL14 group were less than those in the si-NC group. While the invasion cell number and the migration cell number in the LV-METTL14 group were more than those in the LV-NC group, respectively. The differences were statistically significant (all P<0.01). Conclusion:Patients with high METTL14 expression have a worse prognosis in ovarian cancer, which may increase the m6A modification of ovarian cancer cells and promote cells proliferation, invasion and migration.